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IKKε is an IκB kinase participating in the control of NF-κB and type I IFN signal pathways in mammals. However, the function of IKKε in regulating immune response is largely unknown in teleost. Herein, an IKKε homologue named AjIKKε was characterized in Japanese eel (Anguilla japonica). AjIKKε has an N-terminal kinase domain, a ubiquitin-like domain, and a coiled coil-containing domain (CC), which is conserved and similar to its counterpart in mammals. Expression analysis showed that AjIKKε could be up-regulated in kidney, spleen, and particularly in liver under the stimulation of poly I:C, LPS, and Aeromonas hydrophila infection. In vitro, the mRNA levels of AjIKKε were significantly provoked in eel liver cells stimulated by LPS and poly I:C, or the different concentrations of A. hydrophila. The overexpression of AjIKKε could not only induce a significantly higher level of promoter activity of human NF-κB, AP-1, and IFN-β in a dose-dependent manner but also up-regulate the activation of promoters of Japanese eel cRel, AP1, IL6, IFN4, IRF3, and IRF7 in HEK293 cells. RNAi studies showed that after AjIKKε was knocked down, the expression levels of IL1, IL6, TNFα, c-Jun, IFN2, IFN3, MX1, MX2, and IRF3 genes were significantly down-regulated in liver, spleen, and kidney of Japanese eels. In addition, the mutants of AjIKKε-K39A, AjIKKε-S174A, and AjIKKε-ΔCC failed to activate Japanese eel IFN4, IRF3 and human IFN-β promoters in HEK293 cells. Collectively, these results suggest that AjIKKε may function as a positive regulator of NF-κB, MAPK, and IRF3-mediated type I IFN signaling pathways related to immune response evoked by bacterial and viral infection.

In this study, particle-reinforced aluminium matrix composites (PRAMCs) of an Al–Cu–Li alloy were prepared using nano-sized TiB₂+TiC particles. The relationship between TiB₂+TiC nanoparticles and T1 precipitates during the ageing process, as well as the influence of TiC + TiB₂ particles on the growth process of T1 precipitates during the ageing process, were investigated. The grain sizes of the A0 and A1 samples were found to be 249.89 μm and 86.42 μm, respectively. The dislocation density is greater in the deformed A1 sample. The coefficient of thermal expansion (CTE) effect generated by TiC and TiB₂ particles stimulates the precipitation process of T1 precipitates. The A1 alloy reached the peak ageing state in a shorter time and yielded a greater number of T1 precipitates. The precipitation process of T1 precipitate is: SSSS→T1P→T1. The transition from T1P (face-centered cubic, FCC) to T1 (hexagonal close packing, HCP) entails a modification in the order of packing. The recently formed T1P precipitate is attached to the established T1 precipitate and extends along the c-axis through the shared copper-rich layer that has formed on both sides of the mature T1 precipitate. The mechanical properties of sample A1 are optimal at T = 22h, with a yield strength, tensile strength, and elongation of 520 MPa, 553 MPa, and 8.2%, respectively. In comparison to the A0 sample, the yield strength and tensile strength exhibited an increase of 5.05% and 5.13%, respectively.

Three types of 7185+TiB2/TiC aluminum matrix composites with different Zn/Mg ratios were prepared in this work through a water-cooled copper casting process. The effects of Zn/Mg ratio on the microstructure and mechanical properties of 20-mm-thick composites were studied by optical microscopy, electron back scattering diffraction, transmission electron microscopy, and tensile mechanical testing. The results showed that the addition of appropriate TiB2/TiC particles refined the as-cast microstructure of the alloy, and with a decrease in Zn/Mg ratio, the grain size distribution became more uniform, the crystalline phase content gradually increased, and the uniformity of the alloy increased. After aging, as the Zn/Mg ratio decreased, the size of the precipitated phase decreased, and the strengthening effect of the alloy improved. The tensile strengths of the surface layer, 1/4 layer, and center position of the A3 alloy were 558, 568, and 572 MPa, respectively. The strength difference among the different thickness layers was the smallest, and the elongation rate was greater than 12%, resulting in the best uniformity.

Characterization of the mechanical properties plays an essential role in the post-processing and evaluation of the functionality of the additively manufactured metallic parts. A number of studies have been focused on the tensile properties of additively manufactured metals. However, the quasi-static compression test of the additively manufactured 316L blocks with different heat-treatment conditions and scanning strategies seems to be overlooked in the literature. This paper aims to provide a comprehensive study of compressive mechanical response in plastic deformation of SS316L fabricated by selective laser melting (SLM). The mechanical response and microstructures in compressive deformation is analyzed for three printing strategies with 0°-, 90°- and 67.5°- scanning and three heat treatment conditions (450 °C for 3 h, 1100 °C for 1 h with furnace cool and 1100 °C for 1 h with water quenching) for selective laser melted (SLMed) stainless steel 316L in comparison with wrought stainless steel 316L in this work. Alteration of mechanical properties, microstructure evolution and compressive deformation mechanism is studied. Melt pool features are not significantly affected by low-temperature heat treatment (450 °C for 3 h) but fully dissolved through high-temperature heat treatment (1100 °C for 1 h). High-temperature heat treatment provides a higher resistance to compressive plastic deformation for SLMed 316L compared with the low-temperature heat-treated and as-built samples where more twinnings are observed. The compressive plastic deformation mechanism of 90°- and 67.5°-scanning samples is similar, which mainly results from twinning-induced plasticity. For 0°-scanning samples, the strong crystallographic texture is the main cause of anisotropic deformation. Modelling and simulation have been conducted to explain the anisotropic deformation mechanism of the 0°-scanning strategy. Simulation results suggest that the morphology difference of laser-scanning tracks and melt pools, which leads to material flow along the laser scanning direction, explains the anisotropic deformation mechanism.

Abstract Carbon–neutral growth is a crucial long-term climatic aim in the context of global warming. This paper introduces complex network theory and explores its potential application to achieve this goal. Specifically, we investigate the spatial and temporal distribution of nodes and sources in the ecological network, and examine whether a relationship exists between the topological index of network nodes and the landscape pattern index of ecological source areas. We also determine the contribution of nodes to the carbon stock of the entire network by exploring the correlation between the carbon stock of nodes and sources to develop an optimization strategy based on the synergistic effect of node-source carbon enhancement. Finally, we test the effect of network optimization through robustness. Our results show that: (1) The correlation topological feature index analysis reveals that the degree distribution of the node network's topological characteristics becomes dispersed and modular, exhibiting the characteristics of small-world networks according to a large clustering coefficient. The heterogeneity and extent of ecological source landscapes have increased by modularity index but remain distributed and locally fragmented; (2) According to correlation analysis, by enhancing the eccentricity of the node topology, the patch cohesion index (COHESION) of the ecological source site can maximize the contribution of the node to the enhancement of the carbon stock benefits of the source site; (3) According to the tests on the robustness of nodes and edges and the robustness of network links, network stability is improved and carbon sink capacity is enhanced. Simultaneously, the restoration and rejuvenation of ecological space through national ecological construction projects can effectively improve the carbon sink within the organized region, contributing to the carbon neutrality aim. This research gives scientific and quantifiable references for potential ecological construction projects for sustainable cities and the optimization of urban ecological space structure.

Abstract Bone regeneration for large, critical-sized bone defects remains a clinical challenge nowadays. Guided bone regeneration (GBR) is a promising technique for the repair of multiple bone defects, which is widely used in oral and maxillofacial bone defects but is still unsatisfied in the treatment of long bone defects. Here, we successfully fabricated a bilayer mineralized collagen/collagen (MC/Col)-GBR membrane with excellent osteoinductive and barrier function by coating the MC particles prepared via in situ biomimetic mineralization process on one side of a sheet-like pure collagen layer. The aim of the present study was to investigate the physicochemical properties and biological functions of the MC/Col film, and to further evaluate its bone regeneration efficiency in large bone defect repair. Fourier-transform infrared spectra and X-ray diffraction patterns confirmed the presence of both hydroxyapatite and collagen phase in the MC/Col film, as well as the chemical interaction between them. stereo microscope, scanning electron microscopy and atomic force microscope showed the uniform distribution of MC particles in the MC/Col film, resulting in a rougher surface compared to the pure Col film. The quantitative analysis of surface contact angle, light transmittance and tensile strength demonstrated that the MC/Col film have better hydrophilicity, mechanical properties, light-barrier properties, respectively. In vitro macrophage co-culture experiments showed that the MC/Col film can effectively inhibit macrophage proliferation and fusion, reducing fibrous capsule formation. In vivo bone repair assessment of a rabbit critical segmental radial defect proved that the MC/Col film performed better than other groups in promoting bone repair and regeneration due to their unique dual osteoinductive/barrier function. These findings provided evidence that MC/Col film has a great clinical potential for effective bone defect repair. Graphic abstract

Lipid accumulation in ovaries is essential for reproductive success in decapods. Understanding of the mechanism of ovarian lipid accumulation is still in its infancy. In this study, we investigated lipid compositions of ovaries and hepatopancreas throughout vitellogenesis and hemolymph in the late-vitellogenic stage in the mud crab Scylla paramamosain. Total lipid content in ovaries consecutively increased from the early to late stage, and the main accumulated lipids were phosphatidylcholine (PC), alkenylglycerophosphoethanolamine (alkenyl-PE), triacylglycerol (TG), cholesteryl ester, and cholesterol. The lipid composition of hemolymph indicated that PC, alkenyl-PE, diacylglycerol (DG), ether DG, monoacylglycerol, and cholesterol were the main lipids transferred to ovaries from hepatopancreas. In hepatopancreas, the total lipid content appeared to increase during vitellogenesis and the late stage was significantly different from early and mid-stages. TG was the predominant lipid class that slightly decreased from the early to mid-stage, but dramatically increased in the late stage. In contrast, PC and alkenyl-PE significantly increased from the early to mid-stage and remained constantly in relatively low proportion until the late stage. These findings suggested PC and alkenyl-PE in the hepatopancreas likely play critical roles in regulating lipid transfer. Esterified fatty acids in ovaries and hepatopancreas were mainly C16:0, C18:0, 16:1, C18:1, C18:2, C20:4, C20:5, and C22:6, of which highly unsaturated fatty acids (HUFA) primarily occurred in alkenyl-PE and phosphatidylserine (PS) but not PC. In hemolymph, PC was abundant in HUFA besides of saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA), while PS was predominantly composed of SFA and MUFA, implying a transformation of HUFA between phospholipids prior to transfer. These findings highlight the importance of phospholipid metabolism in the hepatopancreas to ovarian lipid accumulation. Our study provides novel insights into the mechanism of ovarian lipid accumulation in decapods.

The blockchain provides a reliable and scalable method for enabling source-tracing functionality in large-scale Internet of Things (IoT) systems. Traditional blockchain-based source tracing applications are generally based on the hypothesis that the raw data collected by each IoT node are credible and consistent, which however may not always be the truth. As no mechanism ensures the reliability of the original data collected from the IoT devices, these data may be accidently screwed up or maliciously tampered with before they are uploaded on-chain. To address this issue, we propose the Multi-dimensional Certificates of Origin (MCO) method to filter out the potentially incredible data-till all the data uploaded to the chain are credible. To achieve this, we devise the Multi-dimensional Information Cross-Verification (MICV) and Multi-source Data Matching Calculation (MDMC) methods. MICV verifies whether a to-be-uploaded datum is consistent or credible, and MDMC determines which data should be discarded and which data should be kept to retain the most likely credible/untampered ones in the circumstance when data inconsistency appears. Large-scale experiments show that our scheme ensures on the credibility of data and off the chain with an affordable overhead.

During the micro-injection molding process, the lifetime of mold inserts is severely limited by high-pressure impact conditions and demolding friction. To investigate the tribological properties and durability of mold insert, nickel-based composite mold inserts have been fabricated by electroforming adding polytetrafluoroethylene (PTFE), molybdenum disulfide (MoS2) and tungsten disulfide (WS2) nanoparticles, respectively. The surface morphology of pure nickel (Ni) and composite mold inserts were characterized. The Ni grain sizes of the composite mold inserts significantly decreased without changing the grain orientation and phase composition. The maximum microhardness of Ni-WS2 mold inserts was increased from 278.2 HV of pure Ni to 456.0 HV, an increase of 1.64 times, while the minimum friction coefficient against Si3N4 ball was reduced to 0.25. In addition, Ni-WS2 mold inserts exhibited excellent tribological properties, and their wear resistances were further investigated at different sliding speeds and external loads. With the increase of sliding speed and load, the self-lubricating property of the composite mold inserts would gradually disappear and the wear rate would increase significantly. The formation of a uniform and dense friction transfer film is the key to reducing the friction coefficient and wear rate. Furthermore, the corrosion resistance of the mold inserts was improved to some extent with the addition of the three nanoparticles. In dynamic electrochemical tests, the corrosion current density of Ni-PTFE mold insert was 3.083 ± 0.228 × 10−6 A/cm2 and the corrosion rate was 0.036 ± 0.003 mm/year, which showed the best corrosion resistance.

The feedback regulatory effects of estrogen (E2) and androgen (T) on the gonadotropin-releasing hormone (GnRH) and gonadotropin (GtH) within the brain–pituitary–gonad (BPG) axis in eels with undeveloped ovaries were investigated through in vivo studies. However, the regulatory role of the BPG axis only became apparent during ovary development in the migratory stage. To further elucidate the direct feedback regulation of the BPG axis, female Anguilla japonica underwent artificial induction of vitellogenesis, and the regulation of BPG axis tissues by GtH (human chorionic gonadotropin, hCG), E2, and T was explored through in vitro exposure. The mRNA expression levels of GnRH (mGnRH), GtH (fshb and lhb), and steroid biosynthesis enzymes (cyp11a1, hsd3b, cyp17a1, and cyp17a2) in the diencephalon, pituitary, and ovary, respectively, were determined. The results showed that the expression level of mGnRH in the diencephalon was significantly downregulated by 0.1 IU/mL hCG but upregulated by both 1 nM E2 and higher concentrations of T, suggesting a direct positive feedback regulation of E2 on mGnRH. In the pituitary, the expression levels of fshb and lhb were upregulated by E2, while fshb was suppressed by T. In the ovaries, the expression of cyp11a1 and hsd3b was upregulated by 1 nM E2, whereas T exposure resulted in an opposite effect. Cyp17a1 mRNA levels did not differ significantly with E2 treatment but were upregulated by 1 nM T. These findings suggest that low concentrations of E2 exhibited positive feedback regulation on all three levels (diencephalon, pituitary, and ovary) of the BPG axis, while T showed weaker and differential feedback regulation in BPG axis tissues. Overall, this study’s results revealed the direct feedback regulation of hCG, E2, and T on the BPG axis in eels, a phylogenetic base of teleosts.

Conclusions: This is the largest study using a healthcare database to indicate the prevalence of hyperuricemia in a health check-up population in an economically developed area of China. The current prevalence among the Chinese health check-up population was substantial, with a higher prevalence in males and in the eastern region. Hyperuricemia and its comorbidities warrant greater attention in the developed areas of China.

AbstractIntroduction To estimate the up-to-date prevalence of chronic kidney disease among the health check-up population in economically developed areas of China using estimated glomerular filtration rate, urinary albumin creatinine ratio, and kidney ultrasound.Methods Healthcare data from 38,093 subjects in 10 megalopolises of China who had an annual health check-up in 2021 were used. The overall and stratified prevalence of chronic kidney disease by sex, age, region and comorbidity group was reported. The association between chronic kidney disease and covariates of demographics, and comorbidities were analyzed in the multivariable-adjusted logistic regression model.Results A total of 3837 CKD cases were detected meeting any of the three CKD diagnostic criteria, with a crude prevalence of 10.1% in the study population. Using one criterion of decreased glomerular filtration rate, albuminuria and kidney structural abnormalities alone detected 204 (5.3%), 3289 (85.7%) and 563 (14.7%) cases, respectively. The addition of kidney ultrasound detected 427 (11.1%) structural abnormality cases without decreased GFR and albuminuria. The most common abnormalities were renal masses, hydronephrosis due to obstruction and congenital anomalies of kidney and urinary tract. Female, older age, low city-tier, hypertension, diabetes, obesity, hypertriglyceridemia as well as early disease stages such as pre-hypertension, impaired fasting glucose and overweight were significantly associated with chronic kidney disease.Conclusion Kidney ultrasound helps to amplify the detection of CKD patients, which is a supplement to kidney function and urine protein.

Major histocompatibility complex (MHC) molecules play a critical role in the immune response of vertebrate animals by presenting foreign antigens to T lymphocytes. In this study, we first cloned and identified a classical and a novel nonclassical MHC I α molecules from Japanese eel (Anguilla japonica), named as AjMHC I-UBA and AjMHC I-L, respectively. The full-length cDNA of AjMHC I-UBA contains an open reading frame (ORF) of 1047 bp encoding a predicted protein of 348 amino acids, while AjMHC I-L 1089 bp encodes 362 amino acids. The multiple alignment of the amino acid sequence showed that AjMHC I-UBA and AjMHC I-L consist of an N-terminal MHC I superfamily domain within α1 and α2 helices, an IgC-MHC I α3 domain, and a transmembrane region close to the C-terminal, which are similar to other fish and mammal species. Molecular polymorphism analysis showed that eight different major alleles of AjMHC I-UBA, named as AjMHC I-UBA*0101～1001, were identified from six Japanese eel individuals. Furthermore, a distinguishing signature of the nonclassical L-lineage genes specific motif “HINMTL”, including an N-glycosylation site (NXS/T), was present in the α3 domain of AjMHC I-L sequences. Although the predicted three-dimensional structures of AjMHC I-UBA and AjMHC I-L are similar to that of human MHC I α, phylogenetic analysis showed that these two protein molecules belong to classical MHC I UBA gene of U-lineage and nonclassical MHC I gene of L-lineage, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that the highest expression of AjMHC I-UBA and AjMHC I-L was found in the intestine and the expression level of AjMHC I-UBA in all of the examined tissues was significantly higher than that of AjMHC I-L. The expressions of AjMHC I-UBA and AjMHC I-L in liver, kidney and spleen were significantly induced following injection with the viral mimic poly I:C, LPS, and Aeromonas hydrophila infection. In vitro, the AjMHC I-UBA and AjMHC I-L transcripts of Japanese eel liver cells were significantly enhanced by the treatment of poly I:C or the stimulation of the high concentration of A. hydrophila. Subcellular localization showed that under natural conditions, AjMHC I-UBA and AjMHC I-L were uniformly distributed in the cytoplasm, and aggregated into spots or flakes in the cytoplasm after the stimulation of poly I:C or LPS. These results collectively suggested that AjMHC I-UBA and AjMHC I-L are important components possibly involved in Japanese eel defense against viral and bacterial infection. Taken together, these findings provide valuable insight into the immune function of MHC class I in the immune system of teleost.

The fragile ecological environment and intensive human activities in the Loess Plateau region of China have led to serious soil erosion and adverse weather disasters. To address these issues, the Chinese government has implemented a series of large-scale ecological restoration projects with significant investments and broad public participation. Throughout this process, the ecosystems have undergone substantial artificial intervention. This study utilizes MODIS data to establish a remote sensing ecological index (RSEI) for evaluating the ecosystem quality of Shanxi Province, located in the eastern Loess Plateau, over the past 20 years (2000–2020). Furthermore, the study analyzed the natural and socio-economic factors that influence the local ecosystem quality using Geodetector analysis. The research findings revealed the following: 1) Over the 20-year period of large-scale ecological restoration projects, the ecosystem quality in Shanxi has significantly improved, as indicated by the average RSEI index increasing from 0.48 to 0.57. 2) The southern part of the province has experienced relatively better ecosystem recovery, while the ecosystem quality in the northern and western regions, particularly along the Yellow River, remains poor. 3) Overall, the major factors influencing ecosystem quality are natural geographical factors such as temperature and slope. However, with rapid urbanization, the negative impacts from socio-economic factors represented by GDP have intensified, particularly in the northern regions. Additionally, this study demonstrated that the geographically weighted regression model can better explain the local variations affecting the RSEI and exhibits a high degree of reliability. The research findings provide scientific and effective references for evaluating the ecosystem quality in the Loess Plateau and implementing ecological restoration projects.

Acquiring high-performance calcium phosphate (CaP) ceramics via 3D printing is critical for their applications in bone regenerative repair. In current study, a complete process study of 3D printed CaP ceramics based on digital light processing (DLP) technology was performed to get the ideal implant for the regenerative repair of calvarial defects. The smaller particle size of initial powder would be more favorable to getting slurry with high solid content and low viscosity. CaP green body with high precision and high curing rate could be fabricated at the exposure energy density and exposure time of 6.20 mJ cm−2 and 2 s, respectively. Moreover, the post-curing treatment could eliminate the interlayer cracks and increase the curing rate from 69.78% to 93.91% of green body. After that, the mechanical strength of CaP ceramics increased by 46.34%. In-vitro cellular experiments showed that 3D printed CaP ceramics had good biocompatibility, and could promote the osteoblastic differentiation of BMSCs. In-vivo rat cranial defect implantation revealed 3D printed CaP ceramics exhibited better osteogenic ability than the commercial ones (BAM®), reaching the close level with the autografts. Overall, this study could provide a closed-loop solution for 3D printed CaP ceramics with high-precision and low-defect to induce bone regeneration.

Dark web vendor identification can be seen as an authorship aliasing problem, aiming to determine whether different accounts on different markets belong to the same real-world vendor, in order to locate cybercriminals involved in dark web market transactions. Existing open-source datasets for dark web marketplaces are outdated and cannot simulate real-world situations, while data labeling methods are difficult and suffer from issues such as inaccurate labeling and limited cross-market research. The problem of identifying vendors’ multiple identities on the dark web involves a large number of categories and a limited number of samples, making it difficult to use traditional multiclass classification models. To address these issues, this paper proposes a metric learning-based method for dark web vendor identification, collecting product data from 21 currently active English dark web marketplaces and using a multi-dimensional feature extraction method based on product titles, descriptions, and images. Using pseudo-labeling technology combined with manual labeling improves data labeling accuracy compared to previous labeling methods. The proposed method uses a Siamese neural network with metric learning to learn the similarity between vendors and achieve the recognition of vendors’ multiple identities. This method achieved better performance with an average F1-score of 0.889 and an accuracy rate of 97.535% on the constructed dataset. The contributions of this paper lie in the proposed method for collecting and labeling data for dark web marketplaces and overcoming the limitations of traditional multiclass classifiers to achieve effective recognition of vendors’ multiple identities.

The landscape is a synthesis of cultural and natural processes in a place that can be designed and maintained to improve and regenerate the natural benefits and services of ecosystems. The Sustainable Sites Initiative has developed a rating system that identifies and advances best practices for landscape architecture. The SITES v2 Rating System was produced through a multi-year iterative process and incentives sustainable landscape practices by using an ecosystem services framework. Opportunities for continuing research on how SITES v2 encourages the creation of a landscape sustainability system and drives more sustainable land development will become apparent as it expands in application, scope, and impact. To illustrate the practice and potential of SITES v2 for sustainable landscape design, this research is primarily a literature review with the specific context of the SITES gold-certified project, Navy Pier in Chicago. The results show that SITES v2 is an important tool for transforming theory into the implementation of ecosystem services and guiding design decisions towards sustainable outcomes. It also promotes the entire project's accountability to a higher standard of best practices and results in a more sustainable site. In the context of contemporary landscape architecture practices, SITES v2 may lead to the transformation of sustainable landscape design. This study can promote the ongoing application of SITES v2 and the creation of a better sustainable landscape through design. It holds the potential to highlight how the methodology on which SITES has been created can be linked with the development of future cities and the management of urban landscapes.

IFNAR1, one of the type I IFN receptors, is crucial to mammalian host defense against viral invasion. However, largely unknown is the immunological role of the fish teleost protein IFNAR1, also known as CRFB5. We have successfully cloned the whole cDNA of the Japanese eel’s (Anguilla japonica) CRFB5a homolog, AjCRFB5a. The two fibronectin-3 domains and the transmembrane region (238–260 aa) of AjCRFB5a are normally present, and it shares a three-dimensional structure with zebrafish, Asian arowana, and humans. According to expression analyses, AjCRFB5a is highly expressed in all tissues found, particularly the liver and intestine. In vivo, Aeromonas hydrophila, LPS, and the viral mimic poly I:C all dramatically increased AjCRFB5a expression in the liver. Japanese eel liver cells were reported to express AjCRFB5a more strongly in vitro after being exposed to Aeromonas hydrophila or being stimulated with poly I: C. The membranes of Japanese eel liver cells contained EGFP-AjCRFB5a proteins, some of which were condensed, according to the results of fluorescence microscopy. Luciferase reporter assays showed that AjCRFB5a overexpression strongly increased the expression of immune-related genes in Japanese eel liver cells, such as IFN1, IFN2, IFN3, IFN4, IRF3, IRF5, and IRF7 of the type I IFN signaling pathway, as well as one of the essential antimicrobial peptides LEAP2, in addition to significantly inducing human IFN-promoter activities in HEK293 cells. Additionally, RNA interference (RNAi) data demonstrated that knocking down AjCRFB5a caused all eight of those genes to drastically lower their expression in Japanese eel liver cells, as well as to variable degrees in the kidney, spleen, liver, and intestine. Our findings together showed that AjCRFB5a participates in the host immune response to bacterial infection by inducing antimicrobial peptides mediated by LEAP2 and favorably modulates host antiviral immune responses by activating IRF3 and IRF7-driven type I IFN signaling pathways.

Organizations can play a significant role in advancing gender equality and environmental sustainability. Increasing the number of women on corporate boards has been offered as a means to increase organizational environmental sustainability. We examined relations between board gender diversity and organizational environmental performance in two studies. The first study examined board gender diversity's relations to corporate environmental sustainability initiatives for 79 largest Turkish organizations. Even though only a small proportion of directors among Turkish organizations are women, the results linked their representation on corporate boards to better environmental performance for all categories of environmental sustainability. The second study examined board gender diversity-corporate environmental performance relations using a sample of 1,776 organizations from 45 countries and representing 8 cultural clusters, greatly expanding the cross-cultural scope of our research. Board gender diversity correlated positively with overall environmental performance (r = 0.26), however there was much variability by country. Gender gap/parity at the national level did not appreciably influence board gender diversity-environmental performance associations. However, there were systematic differences between cultural clusters countries belonged to. Associations were positive and sizable for Anglo, Latin Europe, and Middle Eastern cultural clusters, negligible for Nordic Europe, Eastern Europe, and Confucian Asia cultural clusters, or mostly negligible with great variability for the Southern Asian cultural cluster. Findings highlight the important role that national context and culture play in how women's representation on corporate boards translates or fails to translate into organizational environmental sustainability performance.

Large yellow croaker Larimichthys crocea (Richardson) is an important member in family Sciaenidae, and one of most productive mariculture fishes in China. Fluorescence in situ hybridization is a useful tool for cytogenetics and genomics research. Here, we demonstrated that bacterial artificial chromosome (BAC) clones could be used to identify individual chromosomes in large yellow croaker, and also to study chromosome evolution in the related species. By using BAC paired-end sequencing and sequence alignment, 435 BAC clones were anchored to 24 pseudochromosomes of large yellow croaker genome. Among them, 72 BAC clones with low repeat content were selected and passed PCR test, and then grouped by chromosome for FISH test. As a results, there were 67 BAC clones, 2 to 3 BAC clones per chromosome, generated specific and stable signal at expected position. Then, a dual-color FISH probe cocktail composed of 48 of these BAC clones was formulated and used to hybridize metaphase chromosome spreads, resulting in distinct signal patterns on each chromosome pair, which help to distinguish all chromosomes in the metaphase spreads of large yellow croaker. The chromosome-specific BAC-FISH probes were also applied to a close relative of large yellow croaker, Collichthys lucidus, demonstrating that its Y chromosome originated from the fusion of Chr.1 and Chr.7. Thus, our study provides the first set of chromosome-specific FISH probes in family Sciaenidae, which will play an important role in cytogenetics and genomics research in the family.

Large yellow croaker (Larimichthys crocea) is one of the most commercially important marine fish in southeast China. The white spot disease frequently occurs in large yellow croaker leading to a devastating loss. Given that the pathogen, Cryptocaryon irritans, hardly survive in the low-salinity environment, the low-salinity farming is a potential strategy to defend large yellow croaker. In this study, the water salinity was consecutively decreased from 28 ‰ to 0 ‰, during which the survival rate of L. crocea was stable from 28 ‰ to 4 ‰, while a higher mortality was observed at

Ubiquitination is an essential post-translational modification of proteins and involved in many physiological processes, including sperm differentiation, oocyte maturation, and embryonic development. Among them, the ubiquitin-conjugating enzyme E2 plays a crucial role in gametogenesis. However, the participation of E2 in the regulation of the gonad development of crustaceans is still very limited, especially for oogenesis. In this research, we isolated and identified full-length complementary DNA of ubiquitin-conjugating enzyme E2 from mud crab (Scylla paramamosain) (Sp-Ubc2), detected distribution in different tissues by real-time quantitative PCR, and described expression patterns during oogenesis by in situ hybridization. The results demonstrated that the deduced proteins with a ubiquitin-conjugating (UBC) catalytic domain was clustered with other arthropods. In addition, the Sp-Ubc2 was widely distributed in different tissues of female individuals and kept the highest expression level in the ovary. Moreover, the expression level of Sp-Ubc2 changed significantly during oogenesis and reached peak in secondary and tertiary vitellogenesis stages. At the same time, the Sp-Ubc2 signal in the ovary gradually concentrated in nuclei. These results indicated that Sp-Ubc2 might play important roles in the cell cycle of oogenesis. This study would provide insights into the regulatory mechanism of gonad development in S. paramamosain, and it also enriched the theoretical basis of ubiquitin family genes involved in crustacean gonad development.

PurposeThe purpose of this paper is to alleviate the problem of poor robustness and over-fitting caused by large-scale data in collaborative filtering recommendation algorithms.Design/methodology/approachInterpreting user behavior from the probabilistic perspective of hidden variables is helpful to improve robustness and over-fitting problems. Constructing a recommendation network by variational inference can effectively solve the complex distribution calculation in the probabilistic recommendation model. Based on the aforementioned analysis, this paper uses variational auto-encoder to construct a generating network, which can restore user-rating data to solve the problem of poor robustness and over-fitting caused by large-scale data. Meanwhile, for the existing KL-vanishing problem in the variational inference deep learning model, this paper optimizes the model by the KL annealing and Free Bits methods.FindingsThe effect of the basic model is considerably improved after using the KL annealing or Free Bits method to solve KL vanishing. The proposed models evidently perform worse than competitors on small data sets, such as MovieLens 1 M. By contrast, they have better effects on large data sets such as MovieLens 10 M and MovieLens 20 M.Originality/valueThis paper presents the usage of the variational inference model for collaborative filtering recommendation and introduces the KL annealing and Free Bits methods to improve the basic model effect. Because the variational inference training denotes the probability distribution of the hidden vector, the problem of poor robustness and overfitting is alleviated. When the amount of data is relatively large in the actual application scenario, the probability distribution of the fitted actual data can better represent the user and the item. Therefore, using variational inference for collaborative filtering recommendation is of practical value.

A comparison was made between seasonal and diurnal variations in the Integrated Multi-Satellite Retrievals for Global Precipitation Measurement (IMERG) version 06B Final run and TRMM Multi-Satellite Precipitation Analysis (TMPA) 3B42 version 7 products from April 2014 to March 2019. As for earlier IMERG versions, systematic differences between IMERG version 06B precipitation and TMPA precipitation data were larger over the oceans than over land. Systematic annual mean differences between the IMERG and TMPA data over the oceans were smaller for IMERG version 06B than for earlier IMERG versions, possibly because of updated calibration processes. The mean differences between the IMERG version 06B and TMPA data for tropical oceans were relatively smtropical Pacific for all four seasons were not. The diurnal amplitudes of the IMERG were smaller than those of the TMPA over most continents, and the differences increased with mean diurnal amplitudes. The diurnal amplitudes of the IMERG were larger than those of the TMPA data over the oceans. The differences between the phases of the precipitation diurnal harmonics in the IMERG and TMPA datasets varied widely in all four seasons, but the mean phases were almost the same over both the oceans and the land. The sources of the differences in diurnal precipitation amplitudes in the Bay of Bengal and along the west coast of Central America, which showed large diurnal ranges and rather different diurnal amplitudes, were assessed. Differences in seasonal means caused differences in diurnal amplitudes in the Bay of Bengal, but for Central America, differences in diurnal amplitudes were associated with seasonal mean diurnal amplitudes.

Mycobacterium tuberculosis antimicrobial resistance has been continually reported and is a major public health issue worldwide. Rapid prediction of drug resistance is important for selecting appropriate antibiotic treatments, which significantly increases cure rates. Gene sequencing technology has proven to be a powerful strategy for identifying relevant drug resistance information. This study established a sequencing method and bioinformatics pipeline for resistance gene analysis using an Oxford Nanopore Technologies sequencer. The pipeline was validated by Sanger sequencing and exhibited 100% concordance with the identified variants. Turnaround time for the nanopore sequencing workflow was approximately 12 h, facilitating drug resistance prediction several weeks earlier than that of traditional phenotype drug susceptibility testing. This study produced a customized gene panel assay for rapid bacterial identification via nanopore sequencing, which improves the timeliness of tuberculosis diagnoses and provides a reliable method that may have clinical application.

During the demolding process, the interfacial interaction between the polymer and the metal mold insert will lead to the deformation of the micro-structure, which will directly affect the molding quality and performance of injection molded microfluidic chips. In this study, the demolding quality of micro-channels and micro-mixing structures of polycarbonate (PC), polymethyl methacrylate (PMMA), cyclic olefin copolymer (COC), and polystyrene (PS) microfluidic chips for heavy metal detection were investigated by molding experiments. The experimental results showed that the structures of microfluidic chips could be completely replicated. However, tensile deformation and fracture defects were observed at the edges of the micro-structures after demolding. Compared to the Ni mold insert, the calculation of the relative deviation percentages showed that the width of the micro-channel became larger and the depth became smaller, while the dimensions of the micro-mixing structure changes in the opposite direction. Subsequently, a molecular dynamics (MD) simulation model of polymer/nickel (Ni) mold insert for injection molding was established. The changes of adhesion work, demolding resistance and potential energy during demolding were analyzed. The simulation results showed that the polymer structures had some deformations such as necking, molecular chain stretching and voids under the action of adhesion work and demolding resistance. The difference in the contact area with the mold insert directly brought different interfacial interactions. In addition, the potential energy change of the polymer system could be used to quantitatively characterize the demolding deformation of the structure. Overall, the MD method is able to effectively explain the internal mechanisms of interfacial interactions, leading to the demolding deformation of polymer structures from the molecular/atomic scale.

Lipids are the main energy source for embryonic development in oviparous animals. Prior to the utilization and catabolism, lipids are primarily transported from the yolk sac to embryonic tissues. In the present study, cDNA encoding a circulatory large lipid transfer protein (LLTP) superfamily member, the precursor of large discoidal lipoprotein (dLp) and high-density lipoprotein/β-1,3-glucan-binding protein (HDL-BGBP), named dLp/HDL-BGBP of 14,787 bp in length, was cloned from the mud crab Scylla paramamosain. dLp/HDL-BGBP was predicted to encode a 4,831 amino acids (aa) protein that was the precursor of dLp and HDL-BGBP, which were both detected in hemolymph by liquid chromatography–mass spectrometry (LC-MS/MS) analysis. For the intracellular LLTP, three microsomal triglyceride transfer protein (MTP) cDNAs of 2,905, 2,897, and 3,088 bp in length were cloned from the mud crab and were predicted to encode MTP-A of 881 aa, MTP-B of 889 aa, and MTP-C of 919 aa, respectively, which were different merely in the N-terminal region and shared an identical sequence of 866 aa. During embryonic development, the expression level of dLp/HDL-BGBP consecutively increased from the early appendage formation stage to the eye pigment-formation stage, which indicated that HDL-BGBP is probably the scaffolding protein for yolk lipid. For the MTP gene, MTP-C accounted for ~70% of MTP mRNA from the blastocyst stage to the nauplius stage, as well as the pre-hatching stage; MTP-C and MTP-A expression levels were comparable from the early appendage formation stage to the late eye pigment-formation stage; MTP-A was extremely low in blastocyst and gastrula stages; MTP-B was expressed at a relatively low-level throughout embryo development. The variations in the expression profiles among MTP transcripts suggested that MTP might play roles in the lipid droplet maturation and lipoprotein assembly during embryonic development.

“Immune priming” plays a vital part in the immune system of invertebrates, protecting against recurrent infections by pathogens, and can provide some ideas for the prevention and treatment of invertebrate diseases. Many invertebrates have been demonstrated recently to have immune priming, but the relevant mechanisms are not known. Expression of immune system–related genes in the hemocytes and hepatopancreas of the mud crab (Scylla paramamosain) before and after repeated stimulation with Vibrio parahaemolyticus were analyzed by real-time fluorescence quantitative polymerase chain reaction. Some molecules that may participate in the immune priming of S. paramamosain were screened out, and their possible roles in immune priming were interpreted. Crabs injected first with heat-killed V. parahaemolyticus (HkVp group) or physiologic (0.9%) saline (PS group) were rechallenged at 168 h with live V. parahaemolyticus (HkVp+Vp group and PS+Vp group, respectively). The log-rank test shows a significant difference in survival rate between the HkVp+Vp group and the other groups after the ICH (p < 0.05). Expression of genes involved in the toll-like receptor (TLR) signaling pathway and some antimicrobial peptide genes were detected. By, respectively, comparing gene quantification at different time points in hemocytes and the hepatopancreas, the molecules that may play a part in the early stage of the immune priming of S. paramamosain in the hemocytes are found to be down syndrome cell adhesion molecule (Dscam), Hyastatin, Cactus, Arasin, antilipopolysaccharide factor 3 (ALF3), ALF4, ALF5, and ALF6 as well as later acting molecules, such as Crustin, Dorsal, Pelle, and myeloid differentiation factor 88 (MyD88). The molecules that functioned throughout the entire period are TLR and Spaetzle. In the hepatopancreas, the molecules that may play a part in the early stages of immune priming are Dscam, Hyastatin, Arasin, ALF6, Pelle, Spaetzle, Dorsal and, in the later stage, ALF4. The molecules that functioned throughout the entire period are TLR, Crustin, Cactus, MyD88, ALF3, and ALF5. In summary, the immune function of S. paramamosain is enhanced after it receives the same repetitive stimulation by V. parahaemolyticus, indicating immune priming in S. paramamosain. Our study enriches research on immune priming in invertebrates and lays the foundation for further studies revealing the molecular mechanism of immune priming in crabs.

Mud crab (Scylla paramamosain) is one of the most economically-important marine crabs in China. However, research on mechanisms of reproductive regulation is not sufficient. Vitellogenesis-inhibiting hormone (VIH) is a member of the crustacean hyperglycemia hormones (CHH) family, which plays an essential role in the regulation of gonadal development and maturation in crustaceans, and current studies on the regulation of Vih transcription in crabs are relatively rare. Our previous studies on the transcriptional regulation of mud crab Vih (SpVih) have proved that the binding site of Oct4/Sox9 transcription factor may be the key region for positively regulating the expression of SpVih. In this study, the electrophoretic mobility shift assay (EMSA) experiment confirmed that the nuclear protein extracted from the eyestalk could bind to the key region of SpVih promoter, and these specific bindings were dependent on the presence of Oct4/Sox9 binding sites. Two specific binding complex bands were detected in the supershift group of EMSA supershift experiments by Oct4 and Sox9 antibodies, further confirming the specific recognition of these two transcription factors on the key regulatory region of SpVih. In vitro, Oct4 and Sox9 gene overexpression vectors and SpVih core promoter fragment vector were constructed and co-transfected into HEK293T cells. As a result, SpVih activity increased with the concentration of transcription factors. In vivo, when Oct4 and Sox9 dsRNA were injected into the eyestalks of mud crab, respectively, the expression level of SpVih decreased significantly after interference with Oct4 or Sox9, and the expression level of SpVtg in the ovary and hepatopancreatic increased. Both in vitro and in vivo experiments showed that Oct4 and Sox9 had a positive regulatory effect on SpVih. The GST pull-down experiment was carried out by purified Oct4 and Sox9 proteins, and the results showed that there was an interaction between them. It was speculated that they regulated the expression of SpVih through the interaction.

Myostatin (MSTN) is a negative regulator of muscle growth, which restrains the proliferation and differentiation of myoblasts. To understand the role of two mstn genes of Takifugu bimaculatus, the full-length cDNAs of 1131 bp Tbmstn1 and 1,080 bp Tbmstn2 were obtained from the T. bimaculatus’ genomic database, which encodes 376 and 359 amino acids, respectively. The results of qRT-PCR showed that Tbmstn1 was expressed in the eye, kidney, spleen, skeletal muscle, gill, and brain, and the expression level in the skeletal muscle was extremely significantly higher than in other examined tissues. Tbmstn2 was expressed in the skin, skeletal muscle, gill, and brain, and had the highest expression in the skeletal muscle, followed by expression in the brain. Meanwhile, in different stages of embryonic development, the expression of Tbmstn1 started from the gastrula stage. Its expression in the eye-pigment formation stage and hatching stage was significantly higher than that in other stages. The Tbmstn2 was expressed in all examined embryonic stages with different levels, and the highest expression was detected in the eye-pigment formation stage. These results suggested that Tbmstn1 and Tbmstn2 may involve in the development of skeletal muscle, and Tbmstn2 may be related to the formation of nervous system.

Use of food additives, such as colorants and preservatives, is highly regulated because of their potential health risks to humans. Therefore, it is important to detect these compounds effectively to ensure conformance with industrial standards and to mitigate risk. In this paper, we describe the preparation and performance of an ultrasensitive electrochemiluminescence (ECL) sensor for detecting a key food additive, sunset yellow. The sensor uses graphene quantum dots (GQDs) as the luminescent agent and potassium persulfate as the co-reactant. Strong and sensitive ECL signals are generated in response to trace amounts of added sunset yellow. A detection limit (signal-to-noise ratio = 3) of 7.6 nM and a wide linear range from 2.5 nM to 25 μM are demonstrated. A further advantage of the method is that the luminescent reagents can be recycled, indicating that the method is sustainable, in addition to being simple and highly sensitive.

The application of artificial intelligence in physical education (AIPE) has provided new ways to improve learning and teaching activities in physical classes. However, literature reviews that provide a systematic review and analysis of AIPE are limited. To address this gap, this study provided an overview of AIPE-related empirical research. Specifically, it examined the general state of AIPE, algorithms used for AIPE, and the impact and challenges of AIPE. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses, 130 empirical studies related to AIPE were included in the final synthesis. The findings of this study demonstrated that numerous studies have explored the use of AI technologies to enhance physical education classes and training processes. These technologies have been widely employed in athletic performance analysis, health monitoring, and personalized training. AIPE offered great potential for providing personalized instruction, real-time feedback and assessment, and diverse learning environments. However, the use of AI technology poses challenges, including technical reliability and accuracy, privacy and security issues, as well as technical training and teacher support. These findings provide insights for future research on AIPE.

Receptor-interacting protein 3 (RIP3) has been demonstrated to be a key regulator not only in cell death pathways including apoptosis and necroptosis but also in inflammation and host immune responses. In this study, a RIP3 ortholog named Lc-RIP3 is identified in large yellow croaker (Larimichthys crocea). The open reading frame (ORF) of Lc-RIP3 is 1524 bp long and encodes a protein of 507 amino acids (aa). The deduced Lc-RIP3 protein has an N-terminal kinase domain and a C-terminal RHIM domain, and the genome organization of Lc-RIP3 is conserved in teleosts with 12 exons and 11 introns but is different from that in mammals, which comprises 10 exons and 9 introns. Confocal microscopy revealed that Lc-RIP3 is a cytosolic protein. The expression analysis at the mRNA level indicated that Lc-RIP3 is ubiquitously distributed in various tissues/organs, and could be up-regulated under poly I:C, LPS, PGN, and Pseudomonas plecoglossicida stimulation in vivo. Notably, Lc-RIP3 could induce NF-κB but not IRF3 activation. In addition, Lc-RIP3 co-expression with Lc-TRIF, Lc-MAVS, or Lc-IRF3 significantly abolishes the activation of NF-κB but enhances the induction of IRF3 activity. Moreover, NF-κB activity could be up-regulated when Lc-RIP3 is co-expressed with Lc-RIP1 or Lc-IRF7. These results collectively indicate that Lc-RIP3 acts as an important regulator in host innate immune signaling in teleosts.

The stock market is a dynamical system composed of intricate relationships between financial entities, such as banks, corporations, and institutions. Such a complex interactive system can be represented by the network structure. The underlying mechanism of stock exchange establishes a time-evolving network among companies and individuals, which characterise the correlations of stock prices in the time sequential trades. Here, we develop a novel technique in quantum statistics to analyse the financial market evolution. We commence from heat bath analogy where the normalised Laplacian matrix plays the role of the Hamiltonian operator of the network. The eigenvalues of the Hamiltonian specify energy states of the network. These states are occupied by either indistinguishable bosons or fermions with corresponding Bose-Einstein and Fermi-Dirac statistics. Using the relevant partition functions, we develop the thermodynamic entropy to explore dynamic network characterisations. We conduct the experiments to apply this novel method to identify the significant variance in network structure during the financial crisis. The thermodynamic entropy provides an excellent framework to represent the variations taking place in the stock market.

This study explores and analyzes the specific performance of large language models (LLMs) in instructional design, aiming to unveil their potential strengths and possible weaknesses. Recently, the influence of LLMs has gradually increased in multiple fields, yet exploratory research on their application in education remains relatively scarce. In response to this situation, our research, grounded in pedagogical content knowledge theory, initially formulated an instructional design framework based on mathematical problem chains and corresponding prompt instructions. Subsequently, a comprehensive tool for assessing LLM's instructional design capabilities was developed. Utilizing Generative Pretrained Transformer 4, a high school mathematics teaching plan dataset was generated. Finally, the performance of LLMs in instructional design was evaluated. The evaluation results revealed that the teaching plans generated by LLMs excel in setting instructional objectives, identifying teaching priorities, organizing problem chains and teaching activities, articulating subject content, and selecting methods and strategies. Particularly commendable performance was noted in the modules of statistics and functions. However, there is room for improvement in aspects related to mathematical culture and interdisciplinary assessment, as well as in the geometry and algebra modules. Lastly, this study proposes initiatives, such as LLM prompt-based teacher training and the integration of mathematics-focused LLMs. These suggestions aim to advance personalized instructional design and professional development of teachers, offering educators new insights into the in-depth application of LLMs.

The structure, evolution, and function of heat shock proteins 90 (Hsp90s) have been investigated in great detail in fungi and animals. However, studies on the Hsp90 genes in plants are generally limited. Brachypodium distachyon (L.) P. Beauv., as a model plant for cereal crops, has become a potential biofuel grass. During its long evolution, the Hsp90 gene family in Brachypodium has developed some strategies to cope with adverse environments. How the Hsp90 gene family in Brachypodium evolved in different plant lineages and what its role is in plant responses to drought and salt stresses remains to be elucidated. We used a set of different bioinformatics tools to identify 94 Hsp90 genes from 10 species representing four plant lineages and classified into three subgroups. Eight BdHsp90 genes were detected from B. distachyon. The number of exon-intron structures differed in each subgroup, and the motif analysis revealed that these genes were relatively conservative in each group. The fragments duplication and tandem duplication, which are the prime powers for functional diversity, generally occurred during the duplication of the whole plant genome. Transcriptional analysis of the BdHsp90 genes under salt and drought stress conditions indicated that the expression of these genes was delayed or increased at different stress time points; The expression was more affected in that of Bradi3g39630, Bradi4g06370, and Bradi1g30130. Our findings suggest the involvement of BdHsp90s in plant abiotic stress response, and further consolidate our views on the stress response mechanism of Hsp90 in general.

Heat-shock protein 70 (HSP70) is a molecular chaperone that plays critical roles in cell protein folding and metabolism, which helps to protect cells from unfavorable environmental stress. Haliotis diversicolor is one of the most important economic breeding species in the coastal provinces of south China. To date, the expression and transcriptional regulation of HSP70 in Haliotis diversicolor (HdHSP70) has not been well characterized. In this study, the expression levels of HdHSP70 gene in different tissues and different stress conditions were detected. The results showed that the HdHSP70 gene was ubiquitously expressed in sampled tissues and was the highest in hepatopancreas, followed by hemocytes. In hepatopancreas and hemocytes, the HdHSP70 gene was significantly up-regulated by Vibrio parahaemolyticus infection, thermal stress, and combined stress (Vibrio parahaemolyticus infection and thermal stress combination), indicating that HdHSP70 is involved in the stress response and the regulation of innate immunity. Furthermore, a 2383 bp of 5′-flanking region sequence of the HdHSP70 gene was cloned, and it contains a presumed core promoter region, a CpG island, a (TG)39 simple sequence repeat (SSR), and many potential transcription factor binding sites. The activity of HdHSP70 promoter was evaluated by driving the expression of luciferase gene in HEK293FT cells. A series of experimental results indicated that the core promoter region is located between −189 bp and +46 bp, and high-temperature stress can increase the activity of HdHSP70 promoter. Sequence-consecutive deletions of the luciferase reporter gene in HEK293FT cells revealed two possible promoter activity regions. To further identify the binding site of the key transcription factor in the two regions, two expression vectors with site-directed mutation were constructed. The results showed that the transcriptional activity of NF-1 site-directed mutation was significantly increased (p < 0.05), whereas the transcriptional activity of NF-κB site-directed mutation was significantly reduced. These results suggest that NF-1 and NF-κB may be two important transcription factors that regulate the expression of HdHSP70 gene.

Ubiquitin C-terminal hydrolases (UCHLs) are a subset of deubiquitinating enzymes, and are involved in numerous physiological processes. However, the role of UCHLs during gonad development has not been studied in crustaceans. In this study, we have first cloned and analyzed expression profiling of Sp-uchl3 and Sp-uchl5 genes from mud crab Scylla paramamosain. The full-length cDNA of Sp-uchl3 is of 1804 bp. Its expression level in the ovary was significantly higher than in other tissues (p < 0.01), and during gonadal development, its expression in both O1 and O5 stages was significantly higher than in the other three stages of ovaries (p < 0.05), while in T3 it was higher than in the former two stages of testes (p < 0.05). Meanwhile, the full-length cDNA of Sp-UCHL5 is 1217 bp. The expression level in the ovary was significantly higher than in other tissues (p < 0.01). Its expression in ovaries was higher than in testes during gonadal development (p < 0.05). The expression level in the O5 stage was the highest, followed by the O3 stage in ovarian development, and with no significant difference in the testis development (p > 0.05). These results provide basic data showing the role of Sp-UCHL3 and Sp-UCHL5 in the gonad development of the crab.

Xiphophorus models are important for melanoma, sex determination and differentiation, ovoviviparity and evolution. To gain a global view of the molecular mechanism(s) whereby gene expression may influence sexual dimorphism in Xiphophorus and to develop a database for future studies, we performed a large-scale transcriptome study.The 454-FLX massively parallel DNA sequencing platform was employed to obtain 742,771 and 721,543 reads from 2 normalized cDNA libraries generated from whole adult female and male X. maculatus Jp 163 A, respectively. The reads assembled into 45,538 contigs (here, a "contig" is a set of contiguous sequences), of which, 11,918 shared homology to existing protein sequences. These numbers estimate that the contigs may cover 53% of the total number of Xiphophorus transcriptome. Putative translations were obtained for 11,918 cDNA contigs, of which, 3,049 amino acid sequences contain Pfam domains and 11,064 contigs encode secretory proteins. A total of 3,898 contigs were associated with 2,781 InterPro (IPR) entries and 5,411 contigs with 132 KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways. There were 10,446 contigs annotated with 69,778 gene ontology (GO) terms and the three corresponding organizing principles. Fifty-four potential sex differentially expressed genes have been identified from these contigs. Eight and nine of these contigs were confirmed by real-time PCR as female and male predominantly expressed genes respectively. Based on annotation results, 34 contigs were predicted to be differentially expressed in male and female and 17 of them were also confirmed by real-time PCR.This is the first report of an annotated overview of the transcriptome of X. maculatus and identification of sex differentially expressed genes. These data will be of interest to researchers using the Xiphophorus model. This work also provides an archive for future studies in molecular mechanisms of sexual dimorphism and evolution, and can be used in comparative studies of other fish.